THE HUMAN HEAT-SHOCK PROTEIN FAMILY - EXPRESSION OF A NOVEL HEAT-INDUCIBLE HSP70 (HSP70B') AND ISOLATION OF ITS CDNA AND GENOMIC DNA
THE HUMAN HEAT-SHOCK PROTEIN FAMILY - EXPRESSION OF A NOVEL HEAT-INDUCIBLE HSP70 (HSP70B') AND ISOLATION OF ITS CDNA AND GENOMIC DNA
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DOI：
10.1042/bj2670125
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发表时间：
1990-04-01
影响因子：
4.1
通讯作者：
LIM, L
中科院分区：
文献类型：
--
作者：
LEUNG, TKC;RAJENDRAN, MY;LIM, L
人类热休克蛋白多基因家族包含几种高度保守的蛋白质，它们具有共同的结构和功能特性，但其响应代谢应激的诱导程度有所不同。我们分离并鉴定了一种新的人类 HSP70 cDNA、HSP70B'' cDNA 及其相应的基因序列。 HSP70B'' cDNA 杂交选择了编码比主要应激诱导 HSP70 和组成型表达 HSC70 热休克蛋白更基本的 70 kDa 热休克蛋白的 mRNA，其与其他热休克 70 kDa 蛋白结合 ATP 相同。完整的 HSP70B'' 基因已被测序，并且与主要诱导型 HSP70 基因一样，没有内含子。 HSP70B''基因与HSP70基因有77%的序列相似性，与HSC70 cDNA有70%的相似性，其中3''末端的序列差异最大。 HSP70B'' 基因代表功能基因，如使用特定寡核苷酸的 Northern 印迹分析、使用特定 3'' cDNA 序列的杂交选择翻译和 S1 核酸酶保护实验所表明的。 HSP70 mRNA 在 HeLa 细胞中以低浓度存在，并且在成纤维细胞和 HeLa 细胞中很容易通过热或 CdCl2 处理诱导，与此相反，HSP70B'' mRNA 仅在较高温度下被诱导，并且没有表现出基础表达。诱导模式的差异可能是由于HSP70B''基因启动子区域的特殊特征所致。
The human heat-shock protein multigene family comprises several highly conserved proteins with structural and functional properties in common, but which vary in the extent of their inducibility in response to metabolic stress. We have isolated and characterized a novel human HSP70 cDNA, HSP70B'' cDNA, and its corresponding gene sequence. HSP70B'' cDNA hybrid-selected an mRNA encoding a more basic 70 kDa heat-shock protein than both the major stress-inducible HSP70 and constitutively expressed HSC70 heat-shock proteins, which in common with other heat-shock 70 kDa proteins bound ATP. The complete HSP70B'' gene was sequenced and, like the major inducible HSP70 gene, is devoid of introns. The HSP70B'' gene has 77% sequence similarly to the HSP70 gene and 70% similarity to HSC70 cDNA, with greatest sequence divergence towards the 3''-terminus. The HSP70B'' gene represents a functional gene, as indicated by Northern-blot analysis with specific oligonucleotides, hybrid-selected translation with a specific 3'' cDNA sequence and S1 nuclease protection experiments. In contrast with HSP70 mRNA, which is present at low concentrations in HeLa cells and readily induced by heat or CdCl2 treatment in both fibroblasts and HeLa cells, HSP70B'' mRNA was induced only at higher temperature and showed no basal expression. The differences in pattern of induction may be due to the special features of the promoter region of the HSP70B'' gene.